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Interfering Compounds (interfering + compound)
Selected AbstractsA high-throughput monolithic HPLC method for rapid Vitamin C phenotyping of berry fruitPHYTOCHEMICAL ANALYSIS, Issue 5 2006Paul G. Walker Abstract A rapid method for the quantification of ,ascorbic acid (1) in berry fruit by HPLC with photodiode array detection is presented. ,Ascorbic acid was resolved on a C18 monolithic column with aqueous buffer, after which the column was washed with acetonitrile to remove lipophilic compounds prior to re-equilibration for analysis of the next sample. Using the monolithic column format with high mobile phase flow rates, the entire separation, wash and re-equilibration were achieved in 3 min. With the exception of gooseberry (Ribes uva-crispa), for which an interfering compound co-eluted, concentrations of 1 could be determined in a wide range of berry fruits after extraction in metaphosphoric acid without further sample preparation. Using this extraction method, recoveries of 1 in excess of 85% were achieved. Fruit or juice extracts were stable in 5% metaphosphoric acid for at least 4 h and stability could be extended to longer than 150 h by the addition of the reducing agent tris(2-carboxethyl)phosphine hydrochloride. Following validation, the method was utilised for the phenotyping of fruit in a Scottish Crop Research Institute (SCRI) Ribes nigrum L. breeding population of 300 individuals. An improved extraction method allowed extraction, quantification of 1 and data analysis to be undertaken in less than one working week. Copyright © 2006 John Wiley & Sons, Ltd. [source] Photoelectrocatalytic Oxidation of NADH with Electropolymerized Toluidine Blue OELECTROANALYSIS, Issue 2-3 2007Yusuf Dilgin Abstract A poly(Toluidine Blue O) (poly-TBO) modified electrode was successfully prepared by repeated sweeping the applied potential from ,0.6 to +0.8,V (vs. SCE) on a glassy carbon electrode (GCE) in borate buffer solution at pH,9.1,containing 0.1,M NaNO3 and 0.4,mM Toluidine Blue O (TBO). The poly-TBO modified GCE shows electrocatalytic activity toward NADH oxidation in phosphate buffer solution at pH,7.0, with an overpotential of ca. 350,mV lower than that at the bare electrode. The photoelectrocatalytic oxidation of NADH at this electrode was also successfully investigated by using cyclic voltammetry and amperometry at constant potential. When the modified electrode surface was irradiated with a 250,W halogen lamp, a photoelectrocatalytic effect was observed for NADH oxidation and the current was increased about 2.2 times. The applied potential was selected at +100,mV for amperometric and photoamperometric detection of NADH. A linear calibration graph for NADH was obtained in the range between 1.0×10,5 and 1.0×10,3 M and between 5.0×10,6 and 1.0×10,3 M for amperometric and photoamperometric studies, respectively. The effect of some interfering compounds, such as ascorbic acid and dopamine on the electrocatalytic and photoelectrocatalytic oxidation of NADH was tested. [source] Protein extraction for 2-DE from the lamina of Ecklonia kurome (laminariales): Recalcitrant tissue containing high levels of viscous polysaccharidesELECTROPHORESIS, Issue 3 2008Kouhei Nagai Abstract Extraction of proteins from the tissues of laminarialean algae, i.e. kelp, is difficult due to high levels of nonprotein interfering compounds, mainly viscous polysaccharides. To establish proteomic analysis of kelp species, an ethanol/phenol extraction method was developed and compared to other popular methods. Proteins were extracted with phenol from crude protein powder, obtained by homogenizing the kelp tissues in ice-cold ethanol. The ethanol/phenol method produced high-quality proteins of the highest purity from the lamina of Ecklonia kurome, one of the Japanese dominant laminarialean algae. This method gave well-resolved 1-D SDS-PAGE or 2-DE images with low background and the highest number of bands or spots. In particular, proteins with neutral to basic pI,s were efficiently extracted. Furthermore, 27 spots on the 2-DE gel were extensively identified by MALDI-TOF/TOF analysis. To the best of our knowledge, this is the first report of a protocol for protein extraction from kelp tissues that gives satisfactory 2-D protein profiles. It is expected that the protocol can be applied to other algae tissues or other recalcitrant plant tissues containing high levels of nonprotein interfering compounds. [source] An efficient protein preparation for proteomic analysis of developing cotton fibers by 2-DEELECTROPHORESIS, Issue 22 2006Yuan Yao Abstract Preparation of high-quality proteins from cotton fiber tissues is difficult due to high endogenous levels of polysaccharides, polyphenols, and other interfering compounds. To establish a routine procedure for the application of proteomic analysis to cotton fiber tissues, a new protocol for protein extraction was developed by optimizing a phenol extraction method combined with methanol/ammonium acetate precipitation. The protein extraction for 2-DE was remarkably improved by the combination of chemically and physically modified processes including polyvinylpolypyrrolidone (PVPP) addition, acetone cleaning, and SDS replacement. The protocol gave a higher protein yield and vastly greater resolution and spot intensity. The efficiency of this protocol and its feasibility in fiber proteomic study were demonstrated by comparison of the cotton fiber proteomes at two growth stages. Furthermore, ten protein spots changed significantly were identified by MS/tandem MS and their potential relationships to fiber development were discussed. To the best of our knowledge, this is the first time that a protocol for protein extraction from cotton fiber tissues appears to give satisfactory and reproductive 2-D protein profiles. The protocol is expected to accelerate the process of the proteomic study of cotton fibers and also to be applicable to other recalcitrant plant tissues. [source] Detection of chlorinated quinones using interdigitated electrodes coupled with capillary electrophoresisELECTROPHORESIS, Issue 6 2003Keith B. Male Abstract An array of eight interdigitated microband gold electrodes (IDEs) has been developed together with electrophoretic separation for analysis of chlorinated hydroquinones (ClHQs) and benzoquinones (ClBQs). The IDE chip positioned very close to the separation capillary outlet served as an amplification/detection system without the requirement for frequent "capillary-electrode" alignment. ClHQs, electrophoretically migrating to the IDE surface, were oxidized at +1.1 V by seven electrodes of the array and then detected by the remaining electrode, poised at ,0.1 V. Conversely, ClBQs were detected at +1.1 V by the detecting electrode after having been reduced at the 7 adjacent electrodes poised at ,0.1 V. There was an amplification effect on both the detecting electrode as well as the adjacent electrodes because of the recycle between ClHQs and ClBQs. The detecting "amplification" current response was dependent on the potentials applied, the position of the detecting electrode on the array, the number of adjacent electrodes being used for recycling and the distance between the oxidative and reductive electrodes. Micellar electrokinetic chromatography (MEKC) separation of the analytes was achieved using 30 mM sodium dodecyl sulfate (SDS) with a detection limit in the range of 2,20 ,M. In addition to a facile "capillary-electrode" alignment, the important aspect described here was the capability of detecting through recycling a reduced compound (in the case of ClHQs) at a negative potential to circumvent fouling and electroactive interferences. An appealing feature was also the concurrent oxidation/reduction detection for each compound to ascertain peak assignment, as interfering compounds are less likely to exhibit the same oxidative/reductive characteristics and electrophoretic mobilities as the target analytes. [source] Multidimensional GC coupled to MS for the simultaneous determination of oxygenate compounds and BTEX in gasolineJOURNAL OF SEPARATION SCIENCE, JSS, Issue 4-5 2010Danilo Sciarrone Abstract In the present work, carried out in relation to the European and American Directives on the quality of petrol and diesel fuels, the simultaneous determination of the oxygenate compounds and BTEX in gasoline was achieved through the use of a multidimensional GC (MDGC)/MS system, employing a Deans switch-based transfer system, with an innovative configuration; the latter enabled multiple heart-cut transfers with no hint of retention time shift, a phenomenon that can occur in MDGC, providing the possibility to achieve more then 20 different heart-cuts for the compounds of interest. In this study, 20 selected compounds were quantitatively transferred with 12 heart-cuts, from a first to a secondary column, in order to resolve primary column co-elutions. Analyte quantification and identification was achieved through a fast-scanning quadrupole mass analyzer, operated in full scan mode, in order to evaluate also the interfering compounds transferred together with the compounds of interest. The multidimensional method developed was subjected to validation. All attained data were in excellent correlation with results obtained through the UNI-EN 12177:2000, ASTM D 5580-02 and ASTM D 4815-04 MDGC methods, for the determination of benzene, BTEX and oxygenate compounds in gasoline, respectively. [source] A reliable analytical approach based on gas chromatography coupled to triple quadrupole and time-of-flight mass analyzers for the determination and confirmation of polycyclic aromatic hydrocarbons in complex matrices from aquaculture activitiesRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 13 2009Jaime Nácher-Mestre The potential of gas chromatography coupled to tandem mass spectrometry (GC/MS/MS) with a triple quadrupole analyzer (QqQ) has been investigated for the quantification and reliable identification of sixteen polycyclic aromatic hydrocarbons (PAHs) from the EPA priority list in animal and vegetable samples from aquaculture activities, whose fat content ranged from 5 to 100%. Matrices analyzed included fish fillet, fish feed, fish oil and linseed oil. Combining optimized saponification and solid-phase extraction led to high efficiency in the elimination of interfering compounds, mainly fat, from the extracts. The developed procedure minimized the presence of these interfering compounds in the extracts and provided satisfactory recoveries of PAHs. The excellent sensitivity and selectivity of GC/(QqQ)MS/MS in selected reaction monitoring (SRM) allowed to reach limits of detection at pg/g levels. Two SRM transitions were acquired for each analyte to ensure reliable identification of compounds detected in samples. Confirmation of positive findings was performed by GC coupled to high-resolution time-of-flight mass spectrometry (GC/TOFMS). The accurate mass information provided by GC/TOFMS in full acquisition mode together with its high mass resolution makes it a powerful analytical tool for the unequivocal confirmation of PAHs in the matrices tested. The method developed was applied to the analysis of real-world samples of each matrix studied with the result of detecting and confirming the majority of analytes at the µg/kg level by both QqQ and TOF mass spectrometers. Copyright © 2009 John Wiley & Sons, Ltd. [source] Ion-pairing reversed-phase liquid chromatography/electrospray ionization mass spectrometric analysis of 76 underivatized amino acids of biological interest: a new tool for the diagnosis of inherited disorders of amino acid metabolismRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 12 2005Monique Piraud Seventy-six molecules of biological interest for the diagnosis of inherited disorders of amino acids (AA) metabolism have previously been demonstrated to be detectable in electrospray ionization tandem mass spectrometry (ESI-MS/MS) positive mode without derivatization. Reversed-phase liquid chromatography (RPLC) separation on different C18 columns using various perfluorinated carboxylic acids as ion-pairing agents has been found suitable for coupling with MS/MS, and for the separation of AA. A new procedure was optimized in order to replace the usual ion-exchange chromatographic, post-column ninhydrin derivatization, time-consuming routine method. This procedure allowed an adequate separation of all the molecules from other known interfering compounds, and a throughput of two samples per hour. Quantification limits for each molecule were found to be compatible with their measurement in plasma and urine. We validated the qualitative part of the method by analyzing plasma and urine samples from patients affected with several inherited disorders of AA metabolism. We validated the quantification of 16 AA using their stable isotopes as internal standard. The calibration curves were linear over the range 0,3,mM. The quantitative results obtained with the new method on 105 plasma and 99 urine samples were in good agreement with those obtained by the established routine method. Spiking experiments and precision results were also satisfactory. Copyright © 2005 John Wiley & Sons, Ltd. [source] | ||||||||||